大鼠坐骨神经Wallerian溃变过程中LncRNA的差异表达分析

Analysis of LncRNA expressions and relative functions in Wallerian degeneration after rat sciatic nerve injury

目的:采用分子生物学和生物信息学方法筛选大鼠坐骨神经损伤后Wallerian溃变(Wallerian degeneration,WD)过程中的关键长链非编码RNA(long none coding RNA, LncRNA),并分析该关键LncRNA分子在大鼠坐骨神经损伤过程中的作用。方法:采用Affymetrix大鼠Non Coding RNA芯片,分析大鼠坐骨神经损伤后WD过程中差异表达的LncRNA;通过实时定量聚合酶链式反应验证关键LncRNA的表达;预测关键LncRNA相关基因及功能。结果:基因芯片分析结果显示共有显著差异性LncRNA 708个和mRNA 2 649个,上调LncRNA 283个,下调LncRNA 425个。通过编码—非编码基因共表达分析和预测竞争性内源RNA(competitive endogenous RNA, ceRNA)机制,显示在WD过程中发挥作用的LncRNA,其中上调19个,下调17个。WD早期差异表达的LncRNA靶基因11个,WD后期差异表达的LncRNA靶基因31个。结论:大鼠坐骨神经损伤后,LncRNA显著差异性表达,提示LncRNA在周围神经损伤修复与再生过程中可能发挥了一定作用,为进一步分析LncRNA在神经再生中的作用提供了实验基础。

Objective:To investigate the expression change of LncRNAs during Wallerian degeneration(WD)after rat sciatic nerve injury,screening for key LncRNAs in WD and revealing the regulatory mechanism of these key LncRNA in WD in the repair and regeneration of rat nerve injury.Methods:Screening of differential expressed LncRNAs using gene chip microarrays;Bioinformatics analysis of LncRNAs expression trends,key LncRNAs and relative functions.The data were verified by real-time quantitative PCR.Results:Gene chip analysis revealed the significant difference in the expression changes of 708 LncRNAs and 2649 mRNAs.Among them,there were 283 up-regulated and 425 down-regulated LncRNAs.According to the codingnocoding gene co-expression and ceRNA mechanism analysis,there are 19 up-regulated LncRNAs and 17 down-regulated LncRNAs.There are 11 short-time differential LncRNA target genes and 31 long-term differential LncRNA target genes differentially expressed during WD processes.Conclusion:The differentially expressed LncRNA may play important roles in the process of WD.This study provides a basis of LncRNA on the molecular mechanisms of WD after rat sciatic nerve injury.